Molecular Imaging and Therapy Research Unit, Department of Radiologic Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, Thailand.
Division of Transfusion Science, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, Thailand.
Int J Mol Sci. 2021 Aug 17;22(16):8851. doi: 10.3390/ijms22168851.
Cell-based therapy is a highly promising treatment paradigm in ischemic disease due to its ability to repair tissue when implanted into a damaged site. These therapeutic effects involve a strong paracrine component resulting from the high levels of bioactive molecules secreted in response to the local microenvironment. Therefore, the secreted therapeutic can be modulated by preconditioning the cells during in vitro culturing. Herein, we investigated the potential use of magnetic resonance imaging (MRI) probes, the "iron-quercetin complex" or IronQ, for preconditioning peripheral blood mononuclear cells (PBMCs) to expand proangiogenic cells and enhance their secreted therapeutic factors. PBMCs obtained from healthy donor blood were cultured in the presence of the iron-quercetin complex. Differentiated preconditioning PBMCs were characterized by immunostaining. An enzyme-linked immunosorbent assay was carried out to describe the secreted cytokines. In vitro migration and tubular formation using human umbilical vein endothelial cells (HUVECs) were completed to investigate the proangiogenic efficacy. IronQ significantly increased mononuclear progenitor cell proliferation and differentiation into spindle-shape-like cells, expressing both hematopoietic and stromal cell markers. The expansion increased the number of colony-forming units (CFU-Hill). The conditioned medium obtained from IronQ-treated PBMCs contained high levels of interleukin 8 (IL-8), IL-10, urokinase-type-plasminogen-activator (uPA), matrix metalloproteinases-9 (MMP-9), and tumor necrosis factor-alpha (TNF-α), as well as augmented migration and capillary network formation of HUVECs and fibroblast cells, in vitro. Our study demonstrated that the IronQ-preconditioning PBMC protocol could enhance the angiogenic and reparative potential of non-mobilized PBMCs. This protocol might be used as an adjunctive strategy to improve the efficacy of cell therapy when using PBMCs for ischemic diseases and chronic wounds. However, in vivo assessment is required for further validation.
细胞疗法在缺血性疾病中是一种极具前景的治疗方法，因为它具有在植入受损部位后修复组织的能力。这些治疗效果涉及到一种强烈的旁分泌成分，这是由于生物活性分子在响应局部微环境时的高水平分泌而产生的。因此，可以通过在体外培养过程中对细胞进行预处理来调节分泌的治疗物质。在这里，我们研究了磁共振成像（MRI）探针——“铁-槲皮素复合物”或 IronQ 在预处理外周血单核细胞（PBMC）以扩大促血管生成细胞并增强其分泌的治疗因子中的潜在用途。从健康供体血液中获得的 PBMC 在铁-槲皮素复合物的存在下进行培养。通过免疫染色对分化的预处理 PBMC 进行了特征描述。通过酶联免疫吸附试验（ELISA）来描述分泌的细胞因子。通过使用人脐静脉内皮细胞（HUVEC）进行体外迁移和管状形成实验，以研究促血管生成的效果。IronQ 显著增加了单核祖细胞的增殖，并分化为具有造血和基质细胞标记的纺锤形样细胞。这种扩增增加了集落形成单位（CFU-Hill）的数量。从 IronQ 处理的 PBMC 获得的条件培养基含有高水平的白细胞介素 8（IL-8）、IL-10、尿激酶型纤溶酶原激活物（uPA）、基质金属蛋白酶-9（MMP-9）和肿瘤坏死因子-α（TNF-α），并增强了 HUVEC 和成纤维细胞的迁移和毛细血管网络形成，在体外。我们的研究表明，IronQ 预处理 PBMC 方案可以增强非动员 PBMC 的血管生成和修复潜力。该方案可能作为辅助策略，在使用 PBMC 治疗缺血性疾病和慢性伤口时提高细胞治疗的疗效。然而，需要进行体内评估以进一步验证。
